• Journal of Microbiology and Biotechnology
• /
• 제24권4호
• /
• pp.545-555
• /
• 2014

It has been previously demonstrated that laccases exhibit great potential for use in several industrial and environmental applications. In this paper, two laccase isoenzyme genes, lccB and lccC, were cloned and expressed in Pichia pastoris GS115. The sequence analysis indicated that the lccB and lccC genes consisted of 1,563 and 1,584 bp, and their open reading frames encoded 520 and 527 amino acids, respectively. They had 72.7% degree of identity in nucleotides and 86.7% in amino acids. The expression levels of LccB and LccC were up to 32,479 and 34,231 U/l, respectively. The recombinant laccases were purified by ultrafiltration and $(NH_4)_2SO_4$ precipitation, showing a single band on SDS-PAGE, which had a molecular mass of 58 kDa. The optimal pH and temperature for LccB were 2.0 and $55^{\circ}C$ with 2,2'-azinobis-[ 3-ethylbenzthiazolinesulfonic acid (ABTS) as a substrate, whereas LccC exhibited optimal pH and temperature at 3.0 and $60^{\circ}C$. The apparent kinetic parameters of LccB were 0.43 mM for ABTS with a $V_{max}$ value of 51.28 U/mg, and the Km and $V_{max}$ values for LccC were 0.29 mM and 62.89 U/mg. The recombinant laccases were able to decolorize five types of dyes. Acid Violet 43 (100 g/ml) was completely decolorized by LccB or LccC (2 U/ml), and the decolorization of Reactive Blue KN-R (100 g/ml) was 91.6% by LccC (2 U/ml). Thus, the study characterizes useful laccase isoenzymes from T. versicolor that have the capability of being incorporated into the treatment of similar azo and anthraquinone dyes from dyeing industries.

• 한국약용작물학회지
• /
• 제25권5호
• /
• pp.282-289
• /
• 2017

Background: To investigate the possibility of using Asarum sieboldii as an environment-friendly fumigant for protecting organic cultural heritages, the inhibitory effect of A. sieboldii extract against wood rot fungi on Hanji was examined. Methods and Results: The physical, optical, and morphological properties of Hanji inoculated with Trametes versicolor and Tyromyces palustris, and exposed to the n-hexane fraction of A. sieboldii extract, were measured. The physical properties were expressed as weight loss, zero-span tensile strength and viscosity and the optical properties were depicted by luminance and chromaticity ($L^*$, $a^*$, and $b^*$). The results showed that, the n-hexane fraction of A. sieboldii extract inhibited the growth of fungi on Hanji, and preserved its condition. At a concentration of 25 mg, the n-hexane fraction of A. sieboldii extract maintained zero-span tensile strength, increased viscosity, and restricted discoloration of Hanji. It also was confirmed that the weight of fungi infested Hanji exposed to the extract did not decrease. Scanning electron microscopic images revealed that the spores and hyphae of T. versicolor and T. palustris were not present on Hanji during treatment with > 25 mg of the n-hexane fraction of A. sieboldii extract. Conclusions: These results indicate that the n-hexane fraction of A. sieboldii extract by virtue of its antifungal effectiveness may help in preserving Korean paper cultural heritages, including Hanji.

• 한국식품위생안전성학회지
• /
• 제10권1호
• /
• pp.1-6
• /
• 1995

In order to establish the enzyme linked immunosorbent assay(ELISA) of sterigmatocystin produced by Aspergillus versicolor, we experimented and obtained following results. Two of three rabbits which had been immunized with sterigmatocystin-hemiacetal-BSA produced antibodies against sterigmatocystin at 15 weeks. The produced antibodies were specific for sterigmatocystin and sterigmatocystin-hemiacetal but didn't cross react with other sterigmatocystin analogues in a significant degree. DMF : 4% KC1 (18 : 2) mixed solution was most effective to dissolve sterigmatocystin. For the preparation of sample solution to determine sterigmatocystin by ELISA, sample was extracted with CHC13 and dried, than the dried sample was redissolved with 100 ${mu}ell$ DMF + 4% KC1 mixture. 10~1,000 ng/$m\ell$ level of standard sterigmatocystin could be applied to the established ELISA. When artifically contaminated rice were assayed by the ELISA, the average recovery of sterigmatocystin spiked to 25~500 ng/g was 109% (97~116%), and mean interwell coefficient of variation was 21% (11~28%).

• 한국균학회지
• /
• 제6권1호
• /
• pp.1-4
• /
• 1978

Attempts were made to investigate the sterol components of Coriolus versicolor (Fr.) Quel, which is one of medicinal fungi and which grows wildly in Korea.Its carpophores were collected in the Gyeong Gi Province and extracted with chloroform and methanol. Four Compounds were isolated and one of these compounds was identified as ergosterol by T.L.C., G.L.C. and chemical tests. ${\beta}-Sitosterol$ appears to exist also in the carpophore.

• Biomolecules & Therapeutics
• /
• 제8권3호
• /
• pp.235-240
• /
• 2000

Antitumor effect of LC43, a protein-bound ploysaccharide (M.W. 43 kDa) that was purified from intergeneric protoplast fusant of Lentinus edodes and Coriolus versicolor, was elucidated against mouse sarcoma 180 cell in vitro and in vivo. By injecting LC43 into ICR mice bearing solid or ascitic sarcoma 180, tumor regression and survival rates were investigated. To examine the effects of LC43 on immunopotentiation activity. immunoorgan weight, B cell differentiation, T cell activity and macrophage activation were determined. LC43 showed antitumor effects against both solid tumor and ascitic tumor of sarcoma 180. It did not change significantly the immunoorgan weight but potentiated immune responses such as B cell differentiation and the release of superoxide anion from macrophages. These results suggest that the protein-bound polysaccharide of LC43 exhibited antitumor activities through the activation of immune-related cells and acted as an immunmodulator.

• Journal of Microbiology and Biotechnology
• /
• 제24권12호
• /
• pp.1673-1678
• /
• 2014

The interactive inhibitory effects of pH and chloride on the catalysis of laccase from Trametes versicolor were investigated by studying the alteration of inhibition characteristics of sodium chloride at different pHs for the oxidation of 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid). At pH 3.0, the addition of sodium chloride (50 mM) brought about a 40-fold increase in $K{_m}^{app}$ and a 4-fold decrease in $V_{max}{^{app}}$. As the pH increased to 7.0, the inhibitory effects of sodium chloride became significantly weakened. The mixed-inhibition mechanism was successfully used to quantitatively estimate the competitive and uncompetitive inhibition strengths by chloride at two different pHs (pH 3.0 and 6.0). At pH 3.0, the competitive inhibition constant, $K_i$, was 0.35 mM, whereas the uncompetitive inhibition constant, $K{_i}^{\prime}$, was 18.1 mM, indicating that the major cause of the laccase inhibition by chloride is due to the competitive inhibition step. At a higher pH of 6.0, where the inhibition of the laccase by hydroxide ions takes effect, the inhibition of the laccase by chloride diminished to a great extent, showing increased values of both the competitive inhibition constant ($K_i=23.7mM$) and uncompetitive inhibition constant ($K{_i}^{\prime}=324mM$). These kinetic results evidenced that the hydroxide anion and chloride share a common mechanism to inhibit the laccase activity.

• Mycobiology
• /
• 제39권2호
• /
• pp.103-108
• /
• 2011

Amylases and cellulases are important enzymes that can be utilized for various biological activities. Ten different wild Nigerian mushrooms (Agaricus blazei, Agaricus sp., Corilopsis occidentalis, Coriolus versicolor, Termitomyces clypeatus, Termitomyces globulus, Pleurotus tuber-regium, Podoscypha bolleana, Pogonomyces hydnoides, and Nothopanus hygrophanus) were assayed for production of these secondary metabolites. The results revealed that most of the tested wild fungi demonstrated very good amylase and cellulase activities. With the incorporation of carboxymethyl-cellulose (a carbon source) into the culture medium, Agaricus blazei had the highest amylolytic activity of 0.60 unit/mL (at $25^{\circ}C$, pH 6.8). This was followed in order by P. tuber-regium and Agaricus sp. with 0.42 and 0.39 unit/mL, respectively ($p {\leq} 0.05$). Maltose and sucrose supplementation into the submerged liquid medium made N. hygrophanus and P. hydnoides to exhibit very low amylase activities of 0.09 and 0.11 unit/mL, respectively. Introducing peptone (an organic nitrogen source) into the basal medium enhanced the ability of C. versicolor to produce a cellulase value of 0.74 unit/mL. Other organic nitrogen sources that supported good cellulase activities were yeast extract and urea. Sodium nitrate (inorganic nitrogen source) generally inhibited cellulase production in all mushrooms. The best carbon source was carboxymethyl-cellulose, which promoted very high cellulase activity of 0.67 unit/mL in C. versicolor, which was followed in order by P. tuber-regium, T. chypeatus, and C. occidentalis ($p {\leq} 0.05$). Sucrose was the poorest carbon compound, supporting the lowest values of 0.01, 0.01, and 0.14 unit/mL in P. hydnoides, A. blazei, and Agaricus sp., respectively.

• Journal of Microbiology
• /
• 제45권4호
• /
• pp.333-338
• /
• 2007

Intracellular NADH:quinone reductase involved in degradation of aromatic compounds including lignin was purified and characterized from white rot fungus Trametes versicolor. The activity of quinone reductase was maximal after 3 days of incubation in fungal culture, and the enzyme was purified to homogeneity using ion-exchange, hydrophobic interaction, and gel filtration chromatographies. The purified enzyme has a molecular mass of 41kDa as determined by SDS-PAGE, and exhibits a broad temperature optimum between $20-40^{\circ}C$, with a pH optimum of 6.0. The enzyme preferred FAD as a cofactor and NADH rather than NADPH as an electron donor. Among quinone compounds tested as substrate, menadione showed the highest enzyme activity followed by 1,4-benzoquinone. The enzyme activity was inhibited by $CuSO_4,\;HgCl_2,\;MgSO_4,\;MnSO_4,\;AgNO_3$, dicumarol, KCN, $NaN_3$, and EDTA. Its $K_m\;and\;V_{max}$ with NADH as an electron donor were $23{\mu}M\;and\;101mM/mg$ per min, respectively, and showed a high substrate affinity. Purified quinone reductase could reduce 1,4-benzoquinone to hydroquinone, and induction of this enzyme was higher by 1,4-benzoquinone than those of other quinone compounds.

• Journal of the Korean Wood Science and Technology
• /
• 제31권1호
• /
• pp.1-9
• /
• 2003

항균활성이 우수한 헛개나무 목부 에탄올 조추출물로부터 7종의 화합물을 단리하였으며, 기기분석 결과 flavonoid인 5-hydroxy-7-methoxyflavone를 비롯하여 5,7-dihydroxyflavone (chrysin), 5,7-dihydroxyflavanone (pinocembrin), 3,5,7-trihydroxyflavanone (pinobanksin), 3,4',5,7-tetrahydroxyflavanone (aromadendrin)과 stilbenoid인 3-hydroxy-5-methoxystilbene과 3,5-dihydroxystilbene (pinosylvin)으로 각각 동정되었다. 이들 단리물질에 대한 항균활성을 조사한 결과 stilbenoid인 3-hydroxy-5-methoxystilbene이 공시균주의 생장을 완전히 저해하여 단리물질 중 활성이 가장 높은 것으로 나타났다. 그 다음이 pinocembrin과 pinosylvin으로 Cryphonectria parasitica, Trametes versicolor, Tyromyces palustris 그리고 Trichoderma viride에 대해 높은 균사생장억제율을 나타내어 활성이 우수한 것으로 나타났다. 그러나, flavonoid인 pinobanksin, 5- hydroxy-7-methoxyflavone, chrysin, 그리고 aromadendrin은 항균활성이 낮은 것으로 나타났다. 이상의 결과, 헛개나무 목부 에탄올 조추출물의 높은 항균활성은 3-hydroxy-5-methoxystilbene과 pinocembrin, 그리고 pinosylvin에서 유래된 것으로 사료되었다.

• 자원리싸이클링
• /
• 제18권5호
• /
• pp.52-62
• /
• 2009

두부비지를 이용한 친환경적인 목재방부제를 조제하고, 두부비지 가수분해에 사용하는 산의 농도 및 염의 구성에 따른 방부능의 영향을 조사하는 실험을 수행하였다. 두부비지 방부제는 $25^{\circ}C$에서 1시간 동안 0, 1%, 그리고 2% 황산에 의해 가수분해된 두부비지와 염화구리 또는/그리고 붕산나트륨으로 조제되었고, 목재시편 내에 감가압법으로 주입하여 주입된 시편을 $70^{\circ}C$ 열수에서 72시간 동안의 용탈실험을 거친 후, 갈색부후균 Trametes palustris 및 백색부후균 Tyromyces versicolor에 대한 내후성을 조사하였다. 실험결과 두부비지를 가수분해한 산의 농도가 높아짐에 따라 방부제의 주입능과 내후성이 증가하였고, 용탈성은 감소하였다. 두부비지산 가수분해물/구리염 또는 두부비지 산 가수분해물/구리염/붕소염으로 처리된 목재시편은 T. palustris 및 T. versicolor에 대한 우수한 방부효능을 보였으나, 두부비지 산 가수분해물/붕소염 또는 두부비지가 포함되지 않은 방부제로 처리된 목재시편은 T. palustris에 의한 부후가 관찰되었다. 상기 결과를 토대로 1% 황산에 의해 가수분해한 두부비지, 염화구리 그리고 붕산나트륨으로 조제한 목재방부제가 두부비지를 이용한 목재방부제 조제의 최적 조건으로 평가되었다.